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- EMDB-42048: H3N2 A/Victoria/3/1975 complexed with mouse polyclonal Fab - wk 7... -
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Open data
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Basic information
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Title | H3N2 A/Victoria/3/1975 complexed with mouse polyclonal Fab - wk 7, control immunogen | |||||||||
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![]() | Complex / Fab / Hemagglutinin / polyclonal / IMMUNE SYSTEM | |||||||||
Biological species | ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 17.6 Å | |||||||||
![]() | Carter JJ / Barnes CO | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Vaccine design via antigen reorientation. Authors: Duo Xu / Joshua J Carter / Chunfeng Li / Ashley Utz / Payton A B Weidenbacher / Shaogeng Tang / Mrinmoy Sanyal / Bali Pulendran / Christopher O Barnes / Peter S Kim / ![]() Abstract: A major challenge in creating universal influenza vaccines is to focus immune responses away from the immunodominant, variable head region of hemagglutinin (HA-head) and toward the evolutionarily ...A major challenge in creating universal influenza vaccines is to focus immune responses away from the immunodominant, variable head region of hemagglutinin (HA-head) and toward the evolutionarily conserved stem region (HA-stem). Here we introduce an approach to control antigen orientation via site-specific insertion of aspartate residues that facilitates antigen binding to alum. We demonstrate the generalizability of this approach with antigens from Ebola, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza viruses and observe enhanced neutralizing antibody responses in all cases. We then reorient an H2 HA in an 'upside-down' configuration to increase the exposure and immunogenicity of HA-stem. The reoriented H2 HA (reoH2HA) on alum induced stem-directed antibodies that cross-react with both group 1 and group 2 influenza A subtypes. Electron microscopy polyclonal epitope mapping (EMPEM) revealed that reoH2HA (group 1) elicits cross-reactive antibodies targeting group 2 HA-stems. Our results highlight antigen reorientation as a generalizable approach for designing epitope-focused vaccines. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 437.7 KB | ![]() | |
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Header (meta data) | ![]() ![]() | 14.3 KB 14.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 2.3 KB | Display | ![]() |
Images | ![]() | 18 KB | ||
Filedesc metadata | ![]() | 5.1 KB | ||
Others | ![]() ![]() | 788.9 KB 788.9 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 537.3 KB | Display | ![]() |
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Full document | ![]() | 536.8 KB | Display | |
Data in XML | ![]() | 7.4 KB | Display | |
Data in CIF | ![]() | 9.4 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Voxel size | X=Y=Z: 7.05 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #1
File | emd_42048_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_42048_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
-Entire : H3N2 A/Victoria/3/1975 immune complex with mouse polyclonal Fab -...
Entire | Name: H3N2 A/Victoria/3/1975 immune complex with mouse polyclonal Fab - week 7, control immunogen group |
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Components |
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-Supramolecule #1: H3N2 A/Victoria/3/1975 immune complex with mouse polyclonal Fab -...
Supramolecule | Name: H3N2 A/Victoria/3/1975 immune complex with mouse polyclonal Fab - week 7, control immunogen group type: complex / ID: 1 / Parent: 0 / Macromolecule list: all Details: Recombinantly expressed H3HA with polyclonal Fabs generated from pooled H2HA-immunized mouse antisera |
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-Supramolecule #2: H3N2 A/Victoria/3/1975
Supramolecule | Name: H3N2 A/Victoria/3/1975 / type: complex / ID: 2 / Parent: 1 |
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Source (natural) | Organism: ![]() |
-Supramolecule #3: Polyclonal Fab
Supramolecule | Name: Polyclonal Fab / type: complex / ID: 3 / Parent: 1 |
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Source (natural) | Organism: ![]() ![]() |
-Macromolecule #1: H3N2 A/Victoria/3/1975
Macromolecule | Name: H3N2 A/Victoria/3/1975 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() |
Sequence | String: MKTIIALSYI FCLVFAQDLP GNDNNSTATL CLGHHAVPNG TLVKTITNDQ IEVTNATELV QSSSTGKICN NPHRILDGIN CTLIDALLGD PHCDGFQNEK WDLFVERSKA FSNCFPYDVP DYASLRSLVA SSGTLEFINE GFNWTGVTQN GGSSACKRGP DSGFFSRLNW ...String: MKTIIALSYI FCLVFAQDLP GNDNNSTATL CLGHHAVPNG TLVKTITNDQ IEVTNATELV QSSSTGKICN NPHRILDGIN CTLIDALLGD PHCDGFQNEK WDLFVERSKA FSNCFPYDVP DYASLRSLVA SSGTLEFINE GFNWTGVTQN GGSSACKRGP DSGFFSRLNW LYKSGSTYPV QNVTMPNNDN SDKLYIWGVH HPSTDKEQTN LYVQASGKVT VSTKRSQQTI IPNVGSRPWV RGLSSRISIY WTIVKPGDIL VINSNGNLIA PRGYFKMRTG KSSIMRSDAP IGTCSSECIT PNGSIPNDKP FQNVNKITYG ACPKYVKQNT LKLATGMRNV PEKQTRGIFG AIAGFIENGW EGMIDGWYGF RHQNSEGTGQ AADLKSTQAA IDQINGKLNR VIEKTNEKFH QIEKEFSEVE GRIQDLEKYV EDTKIDLWSY NAELLVALEN QHTIDLTDSE MNKLFEKTRR QLRENAEDMG NGCFKIYHKC DNACIGSIRN GTYDHDVYRD EALNNRFQIK GSMKQIEDKI EEILSKIYHI ENEIARIKKL IGEVASSSGL NDIFEAQKIE WHEAHHHHHH G |
-Experimental details
-Structure determination
Method | negative staining |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.015 mg/mL |
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Buffer | pH: 7 |
Staining | Type: NEGATIVE / Material: Uranyl Formate |
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Electron microscopy
Microscope | TFS GLACIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 15.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm |